LOCUS       sequence-314104-        7045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human PLIN3 with a N-terminal GFP-FLAG tag. Confers 
            Puromycin resistance. Inducible in T-REx cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7045)
  AUTHORS   Demange P, Joly E, Marcoux J, Zanon PRA, Listunov D, Rulliere P, 
            Barthes C, Noirot C, Izquierdo JB, Rozie A, Pradines K, Hee R, de 
            Brito MV, Marcellin M, Serre RF, Bouchez O, Burlet-Schiltz O, 
            Oliveira MCF, Ballereau S, Bernardes-Genisson V, Maraval V, Calsou 
            P, Hacker SM, Genisson Y, Chauvin R, Britton S
  TITLE     SDR enzymes oxidize specific lipidic alkynylcarbinols into cytotoxic
            protein-reactive species.
  JOURNAL   Elife. 2022 May 10;11. pii: 73913. doi: 10.7554/eLife.73913.
  PUBMED    35535493
REFERENCE   2  (bases 1 to 7045)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2022-05-10"; volume: "11. pii: 73913. doi"; pages: " 
            10.7554/eLife.73913"
COMMENT     Sequence Label: pICE-EGFP-FLAG-PLIN3
FEATURES             Location/Qualifiers
     source          1..7045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             733..756
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2107..2124)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2113..2337
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2383..2811
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2470..2489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2680..2701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2820..2840)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2825..3154
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3005..3140
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3067..3086
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3206..3805
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3206..3225)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3702..3722
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3935..4056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3972..3991)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4026..4045
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4106..4122)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4106..4122)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4119..4141)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4130..4146
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4154..4160,4161..4178,4179..4184))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4199..4220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4337..4354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4508..5093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4588..4607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5264..6055,6056..6124))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5887..5906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6125..6229)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6301..6680
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6681..6884
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6835..6855
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6886..6904
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6907..6925
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6935..6959
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      join(7040..7045,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc
      721 ggactcagat ctgattacaa ggatgacgac gataagctcg agtctgccga cggggcagag
      781 gctgatggca gcacccaggt gacagtggaa gaaccggtac agcagcccag tgtggtggac
      841 cgtgtggcca gcatgcctct gatcagctcc acctgcgaca tggtgtccgc agcctatgcc
      901 tccaccaagg agagctaccc gcacatcaag actgtctgcg acgcagcaga gaagggagtg
      961 aggaccctca cggcggctgc tgtcagcggg gctcagccga tcctctccaa gctggagccc
     1021 cagattgcat cagccagcga atacgcccac agggggctgg acaagttgga ggagaacctc
     1081 cccatcctgc agcagcccac ggagaaggtc ctggcggaca ccaaggagct tgtgtcgtct
     1141 aaggtgtcgg gggcccaaga gatggtgtct agcgccaagg acacggtggc cacccaattg
     1201 tcggaggcgg tggacgcgac ccgcggtgct gtgcagagcg gcgtggacaa gacaaagtcc
     1261 gtagtgaccg gcggcgtcca atcagtcatg ggctcccgct tgggccagat ggtgctgagt
     1321 ggggtcgaca cggtgctggg gaagtcggag gagtgggcgg acaaccacct gccccttacg
     1381 gatgccgaac tggcccgcat cgccacatcc ctggatggct tcgacgtcgc gtccgtgcag
     1441 cagcagcggc aggaacagag ctacttcgta cgtctgggct ccctgtcgga gaggctgcgg
     1501 cagcacgcct atgagcactc gctgggcaag cttcgagcca ccaagcagag ggcacaggag
     1561 gctctgctgc agctgtcgca ggccctaagc ctgatggaaa ctgtcaagca aggcgttgat
     1621 cagaagctgg tggaaggcca ggagaagctg caccagatgt ggctcagctg gaaccagaag
     1681 cagctccagg gccccgagaa ggagccgccc aagccagagc aggtcgagtc ccgggcgctc
     1741 accatgttcc gggacattgc ccagcaactg caggccacct gtacctccct ggggtccagc
     1801 attcagggcc tccccaccaa tgtgaaggac caggtgcagc aggcccgccg ccaggtggag
     1861 gacctccagg ccacgttttc cagcatccac tccttccagg acctgtccag cagcattctg
     1921 gcccagagcc gtgagcgtgt cgccagcgcc cgcgaggccc tggaccacat ggtggaatat
     1981 gtggcccaga acacacctgt cacgtggctc gtgggaccct ttgcccctgg aatcactgag
     2041 aaagccccgg aggagaagaa gtaggcggcc gcacgcgtct gcagtctaga taaacccgct
     2101 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2161 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2221 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2281 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2341 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2401 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2461 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2521 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2581 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2641 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2701 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2761 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2821 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2881 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2941 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3001 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3061 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3121 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3181 tttcggatct gatcagcacg tgaccatgac cgagtacaag cccacggtgc gcctcgccac
     3241 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     3301 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     3361 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     3421 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     3481 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     3541 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     3601 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     3661 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     3721 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     3781 catgacccgc aagcccggtg cctgacacgt gctacgagat ttcgattcca ccgccgcctt
     3841 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3901 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     3961 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4021 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctag ccgtcgacct
     4081 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4141 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4201 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4261 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4321 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4381 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4441 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4501 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4561 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4621 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4681 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4741 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4801 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4861 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4921 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4981 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5041 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5101 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5161 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5221 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5281 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5341 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5401 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5461 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5521 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5581 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5641 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5701 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5761 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5821 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5881 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5941 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6001 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6061 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6121 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6181 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6241 gaaaagtgcc acctgacgtc gacggatcgg gagatcctgt acgggccaga tatccgcgtt
     6301 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     6361 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     6421 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     6481 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     6541 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     6601 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     6661 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     6721 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     6781 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     6841 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
     6901 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
     6961 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
     7021 tccggcaagc ttaccggtcg ccacc
//
